Characterization and Biocompatibility of Exosomes from Human Amniotic Membrane-Derived Mesenchymal Stem Cells in Tissue Engineering and Regenerative Medicine: The Impact of Hypoxia Condition

Background: Hypoxia is a known method to stimulate stem cells and enhance exosome production, potentially improving their therapeutic qualities. This study investigates how hypoxia affects the biological behavior of human amniotic membrane-derived mesenchymal stem cells (AM-MSCs) and the structural and functional properties of their exosomes. Methods: AM-MSCs were isolated from healthy placentas and cultured under normoxic (۲۲% O₂) or hypoxic (۱% O₂) conditions. Cell count, viability, apoptosis, migration, and differentiation potential into osteocytes, adipocytes, and chondrocytes were assessed. Exosomes were then isolated and analyzed for morphology (size, shape, distribution), structure (aggregation, colloidal behavior, zeta potential), protein concentration, and biocompatibility. Results: Hypoxia (۱% O₂) did not adversely affect AM-MSC viability, apoptosis, or differentiation (P > ۰.۰۵) but significantly enhanced cell migration (P ≤ ۰.۰۰۱). Exosomes from hypoxia-treated cells had smaller size (۱۴۵.۴ nm vs. ۱۸۵.۷ nm), higher uniformity (PI = ۰.۴۲۰ vs. ۰.۷۵۶), and better stability (zeta potential: -۲.۳۷ mV vs. -۱۲.۵۷ mV). They also contained more total protein (۵.۰۰۳ mg/ml vs. ۴.۱۰۹ mg/ml) and showed no toxicity in biocompatibility tests. Conclusion: Hypoxic preconditioning enhances the migration capacity of AM-MSCs and improves the structural and biochemical profile of their exosomes, suggesting a superior potential for therapeutic use in cell-free treatments.

کلیدواژه ها:

Exosome ، Hypoxia ، Mesenchymal stem cell ، Amniotic membrane ، Migration